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Il17A autoantibody induced by recombinant protein Ag85A-Il17A

Applied Biochemistry and Biotechnology, ISSN: 0273-2289, Vol: 169, Issue: 2, Page: 502-510
2013
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Article Description

IL17A is a widely studied cytokine which plays an important role in autoimmune diseases as well as asthma. In this study, murine IL17a gene fragment was inserted into pET28a-Ag85A and it was expressed as a fusion protein named Ag85A-IL17A. Recombinant protein Ag85A-IL17A was expressed as an open reading frame of 1,287 bp, encoding a polypeptide of 429 amino acid residues with a predicted molecular mass of 50 kDa. It was expressed in Escherichia coli. BL21 and purified by metal affinity chromatography using a nickel-nitrilotriacetic acid column. Bioinformatics revealed that Ag85A-IL17A contained B cell epitopes of IL17A. Purified proteins Ag85A-IL17A and Ag85A were injected with adjuvants into mice then antibody responses in sera were measured by enzyme-linked immunosorbent assay and western blot assay. Experiments showed that purified protein Ag85-IL17A could bind to a standard IL17A antibody and could induce IL17A autoantibody in mice. © 2012 Springer Science+Business Media New York.

Bibliographic Details

Jin, Rong; Guo, Sheng; Wu, Liangxia; Fan, Xiaoyong; Ma, Hui; Lowrie, Douglas B; Zhang, Jianhua

Springer Science and Business Media LLC

Biochemistry, Genetics and Molecular Biology; Chemical Engineering; Immunology and Microbiology

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